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Collagen cross-linked with human stem cell factors, a preparation method of collagen and applications of collagen in fields of beauty treatment and skin repair

A technology of stem cell factor and collagen, which is applied in the field of preparation and cross-linking collagen of human stem cell factor to achieve the effect of repairing skin wounds

Active Publication Date: 2020-04-10
SHANDONG XINRUI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Stem cells and stem cell factors have proved their effectiveness in the treatment of many diseases, but they still encounter many clinical application problems, one of which is how to ensure that stem cells and their secreted cytokines work at the site of disease occurrence, and It will not be wasted in normal parts or non-treatment demand parts, which will affect the final treatment results

Method used

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  • Collagen cross-linked with human stem cell factors, a preparation method of collagen and applications of collagen in fields of beauty treatment and skin repair
  • Collagen cross-linked with human stem cell factors, a preparation method of collagen and applications of collagen in fields of beauty treatment and skin repair
  • Collagen cross-linked with human stem cell factors, a preparation method of collagen and applications of collagen in fields of beauty treatment and skin repair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Obtaining Human Stem Cell Factors with Collagen Affinity

[0038] Coat the bottom of the cell culture flask with collagen, coating method: 10mg collagen I, 99.167mL H 2 0.0.833mL 6M HCl is used for reagent preparation, and after standing at room temperature for 3 hours, use a 0.45um filter to filter, and the plating concentration in the cell bottle is 0.5ml / cm 2 , the bottom is naturally dry, and the coated culture bottle or culture plate can be stored at a low temperature of 4 degrees for one year.

[0039] Obtain P3 umbilical cord mesenchymal stem cells from the cell bank, inoculate them in culture flasks coated with collagen type I, and press 1-3×10 4 / cm 2 For inoculation, place at 37 °C, 5% CO 2 Culture in the incubator for 2-3 days, and when the cell confluency reaches 85-90%, the cell supernatant is collected.

Embodiment 2

[0041] Human Stem Cell Factor Concentrate

[0042] The collected stem cell culture supernatant was filtered through a 0.22 μm filter membrane, and then passed through a Vivaflow 50 swirl / tangential flow ultrafilter with a molecular weight of 50KD to concentrate the stem cell supernatant, and then the concentrated solution was passed through a Vivaflow with a molecular weight of 3KD 50 swirl flow / tangential flow ultrafilter to collect the retentate, which is the cytokine concentrate.

Embodiment 3

[0044] ELISA detection of concentrated human stem cell factor concentration

[0045]ELISA quantitative kits were used to detect the contents of SCF, VEGF, IL-10, and TGF-β in stem cell factors respectively. Taking TGF-β as an example, the detailed steps of ELISA kit determination were described.

[0046] (1) Mix all reagents thoroughly before use. Do not make the liquid produce a lot of foam, so as not to add a lot of bubbles when adding the sample, resulting in errors in the sample addition.

[0047] (2) Determine the number of strips required based on the number of samples to be tested plus the number of standards. Each standard, blank well, and sample were repeated three times. The sample was diluted 1:1 with the sample diluent and added 50uL to the reaction well.

[0048] (3) Add 50uL of the diluted standard substance to the reaction well, and add 50uL of the sample to be tested into the reaction well. Immediately add 50uL of biotinylated antibody. Cover the membrane ...

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PUM

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Abstract

The invention discloses collagen cross-linked with human stem cell factors, which comprises collagen obtained by cross-linking and curing the human stem cell factors with collagen affinity and collagen by using a cross-linking agent. The invention also discloses a preparation method of the collagen cross-linked with human stem cell factors and applications of the collagen in the fields of beauty treatment and skin repair. The collagen can promote wound healing, improve skin quality and skin laxity, reduce skin wrinkles and stripes, improve skin compactness, whiten skin, reduce skin darkness and skin spots and remove acnes.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to collagen cross-linked with human stem cell factors, a preparation method and application thereof. Background technique [0002] Stem cells are undifferentiated cells with the ability to self-replicate and differentiate into various functional cells. Stem cell factor is a group of important components in the microenvironment of stem cell survival. It has been proved to play an important role in the treatment of human immune diseases, tissue wound repair and anti-aging by stem cells. Stem cells from different sources can secrete different groups of stem cell factors, and the amount of stem cell factors secreted by the same type of stem cells in different culture systems will also be different. If umbilical cord-derived mesenchymal stem cells are used for in vitro culture, a large amount of stem cell factors with low immunogenicity can be obtained. Stem cells and stem c...

Claims

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Application Information

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IPC IPC(8): A61K47/64A61K38/19A61K8/65A61K8/64A61P17/02A61P17/16A61P17/18A61P37/02A61Q19/00A61Q19/02A61Q19/08C12P21/02C07K14/52C07K1/34
CPCA61K47/6435C12P21/02C07K14/52A61K38/19A61K8/65A61K8/64A61Q19/00A61Q19/08A61Q19/02A61P17/18A61P17/02A61P37/02A61P17/16
Inventor 刘明录王立新强邦明张传鹏韩庆梅金海锋冯建海王翠莲刘媛媛
Owner SHANDONG XINRUI BIOTECH CO LTD